Production, characterisation and applications of monoclonal antibodies to two novel porcine bocaviruses from swine in Northern Ireland

The production, preliminary characterisation and applications of monoclonal antibodies (mAbs) against two novel swine bocaviruses isolated in cell culture from swine in Northern Ireland are described. Of the 17 stable final clones produced, four were characterised. All were of the IgG2a isotype and showed no cross-reactivity with either bocavirus strain. Partial neutralisation was observed with PBoV4 mAbs and homologous virus. The two mAbs selected for use in antigen-detecting ELISAs were successful in highlighting those fractions containing infectious virus within sucrose gradients. This is the first report of the production of specific reagents that will prove useful in the study of the biology of these viruses and swine bocavirus-associated diseases.

Isolation in cell cultures and initial characterisation of two novel bocavirus species from swine in Northern Ireland

We report the isolation in cell cultures of two novel bocavirus species in pigs from farms in Northern Ireland with clinical postweaning multisystemic wasting syndrome (PMWS). We have designated the isolates as porcine bocavirus-3 (PBoV3) and porcine bocavirus-4 (PBoV4). To date 5082 and 4125 bps of PBoV3 and PBoV4 have been sequenced, respectively. PBoV3 and PBoV4 show nucleotide homology to other known bocaviruses in swine and other organisms. Open reading frame (ORF) analysis has shown that these viruses have a third small ORF, equivalent to the NP1 ORF that distinguishes the bocaviruses from other parvoviruses.

Development of a minor groove binder assay for real-time one-step RT-PCR detection of swine vesicular disease virus

The design and development of a 5' conjugated minor groove binder (MGB) probe real-time RT-PCR assay are described for rapid, sensitive and specific detection of swine vesicular disease virus (SVDV) RNA. The assay is designed to target the 2C gene of the SVDV genome and is capable of detecting 2 x 10(2) copies of an RNA standard per reaction. It does not detect any of the other RNA viruses that cause vesicular disease in pigs, or the human enterovirus, Coxsackie B5 virus (CVB5) which is closely related antigenically to SVDV. The linear range of this test was from 2 x 10(2) to 2 x 10(8) copies/mu l. The assay is rapid and can detect SVDV RNA in just over 3.5 h including the time required for nucleic acid extraction. The development of this assay provides a useful tool for the differential diagnosis of SVD or for the detection of SVDV in research applications. This study demonstrates the suitability of MGB probes as a real-time PCR chemistry for the diagnosis of swine vesicular disease. (C) 2010 Elsevier B.V. All rights reserved.

Sensitive detection of African swine fever virus using real-time PCR with a 5 ' conjugated minor groove binder probe

The design of a 5' conjugated minor groove binder (MGB) probe real-time PCR assay is described for the rapid, sensitive and specific detection of African swine fever virus (ASFV) DNA. The assay is designed against the 9GL region and is capable of detecting 20 copies of a DNA standard. It does not detect any of the other common swine DNA viruses tested in this study. The assay can detect ASFV DNA in a range of clinical samples. Sensitivity was equivalent to the Office International des Epizooties (OIE) recommended TaqMan assay. In addition the assay was found to have a detection limit 10-fold more sensitive than the conventional PCR recommended by the OIE. Linear range was ten logs from 2 x 10(1) to 2 x 10(10). The assay is rapid with an amplification time just over 2 h. The development of this assay provides a useful tool for the specific diagnosis of ASF in statutory or emergency testing programs or for the detection of ASFV DNA in research applications. (C) 2010 Elsevier B.V. All rights reserved.

Scaled Heavy-Ball Acceleration of the Richardson-Lucy Algorithm

The Richardson-Lucy algorithm is one of the most important algorithms in the image deconvolution area. However, one of its drawbacks is slow convergence. A very significant acceleration is obtained by the technique proposed by Biggs and Andrews (BA), which is implemented in the deconvlucy function of the Image Processing MATLAB toolbox. The BA method was developed heuristically with no proof of convergence. In this paper, we introduce the Heavy-Ball (H-B) method for Poisson data optimization and extend it to a scaled H-B method, which includes the BA method as a special case. The method has proof of the convergence rate of O(k-2), where k is the number of iterations. We demonstrate the superior convergence performance of the scaled H-B method on both synthetic and real 3D images.

Hand-held voltammetric analyser for real-time monitoring of heavy metals in situ

This paper presents a novel hand-held instrument capable of real-time in situ detection and identification of heavy metals. The proposed system provides the facilities found in a traditional lab-based instrument in a hand held a design. In contrast to existing commercial systems, it can stand alone without the need of an associated computer. The electrochemical instrument uses anodic stripping voltammetry which is a precise and sensitive analytical method with excellent limits of detection. The sensors comprise disposable screen-printed (solid working) electrodes rather than the more common hanging mercury drop electrodes. The system is reliable, easy to use, safe, avoids expensive and time-consuming procedures and may be used in a variety of situations to help in the fields of environmental assessment and control.

Environmental forensic investigations: The potential use of a novel heavy metal sensor and novel taggants

This chapter presents a novel hand-held instrument capable of real-time in situ detection and identification of heavy metals, along with the potential use of novel taggants in environmental forensic investigations. The proposed system provides the facilities found in a traditional laboratory-based instrument but in a hand held design, without the need for an associated computer. The electrochemical instrument uses anodic stripping voltammetry, which is a precise and sensitive analytical method with excellent limits of detection. The sensors comprise a small disposable plastic strip of screen-printed electrodes rather than the more common glassy carbon disc and gold electrodes. The system is designed for use by a surveyor on site, allowing them to locate hotspots, thus avoiding the expense and time delay of prior laboratory analysis. This is particularly important in environmental forensic analysis when a site may have been released back to the owner and samples could be compromised on return visits. The system can be used in a variety of situations in environmental assessments, the data acquired from which provide a metals fingerprint suitable for input to a database. The proposed novel taggant tracers, based on narrow-band atomic fluorescence, are under development for potential deployment as forensic environmental tracers. The use of discrete fluorescent species in an environmentally stable host has been investigated to replace existing toxic, broadband molecular dye tracers. The narrow band emission signals offer the potential for tracing a large number of signals in the same environment. This will give increased data accuracy and allow multiple source environmental monitoring of environmental parameters.

Voltammetric in situ measurements of heavy metals in soil using a portable electrochemical instrument

This paper presents a portable electrochemical instrument capable of detecting and identifying heavy metals in soil, in situ. The instrument has been developed for use in a variety of situations to facilitate contaminated land surveys, avoiding expensive and time-consuming procedures. The system uses differential pulse anodic stripping voltammetry which is a precise and sensitive analytical method with excellent limits of detection. The identification of metals is based on a statistical microprocessor-based method. The instrument is capable of detecting six different toxic metals (lead, cadmium, zinc, nickel, mercury and copper) with good sensitivity

Acidity compensation of electrochemical measurements for on-site monitoring of heavy metals

This paper presents an electrochemical instrumentation system capable of real-time in situ detection of heavy metals. A practical approach to introduce acidity compensation against changes in amplitude of the peak currents is also presented. The compensated amplitudes can then be used to predict the concentration level of heavy metals. The system uses differential pulse anodic stripping voltammetry, which is a precise and sensitive analytical method with excellent limits of detection. The instrument is capable of detecting lead, cadmium, zinc, nickel and copper with good sensitivity and precision. The system avoids expensive and time-consuming procedures and may be used in a variety of situations to help environmental assessment and control. 

On-site monitoring and cartographical mapping of heavy metals

This paper presents a portable electrochemical instrument capable of real-time in situ detection and automatic identification of heavy metals. The instrument is equipped with an embedded Geographical Position System and is capable of storing the geographical position of the sample under test. Software has been developed to combine pollutant results with geographical position, in order to produce a cartographical presentation of the pollution of an area. The electrochemical instrument provides the facilities found in a traditional lab based instrument in a portable design for on-site measurements. The instrument is capable of detecting lead, cadmium, zinc, nickel, mercury, and copper with good sensitivity and precision. The system is reliable, easy to use, safe, and it may be used in a variety of situations to help environmental assessment and control.